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Biomedical subjects

David G Lloyd

Publications and source records attributed to David G Lloyd.

At least 19 recordsLinked to original sources

Repeatability of 3D gait kinematics obtained from an electromagnetic tracking system during treadmill locomotion.

The purpose of this paper was to describe a technique that enables three-dimensional (3D) gait kinematics to be obtained using an electromagnetic tracking system, and to report the intra-trial, intra-day/inter-tester and inter-day/intra-tester repeatability of kinematic gait data obtained using this technique. Ten able-bodied adults underwent four gait assessments; the same two testers tested each subject independently on two different days. Gait assessments were conducted on a custom-built long-bed treadmill with no metal components between the rollers. Each gait assessment involved familiarisation to treadmill walking, subject anatomical and functional calibration, and a period of steady-state treadmill walking at a self-selected speed. Following data collection, 3D joint kinematics were calculated using the joint coordinate system approach. 3D joint angle waveforms for 10 left and right strides were extracted and temporally normalised for each trial. Intra-trial, intra-day/inter-tester and inter-day/intra-tester repeatability of the temporally normalised kinematic waveforms were quantified using the coefficient of multiple determination (CMD). CMDs for joint kinematics averaged 0.942 intra-trial, 0.849 intra-day/inter-tester and 0.773 inter-day/intra-tester. In general, sagittal plane kinematics were more repeatable than frontal or transverse plane kinematics, and kinematics at the hip were more repeatable than at the knee or ankle. The level of repeatability of kinematic gait data obtained during treadmill walking using this protocol was equal or superior to that reported previously for overground walking using image-based protocols.

Adolescent↗

Permuting input for more effective sampling of 3D conformer space.

SMILES strings and other classic 2D structural formats offer a convenient way to represent molecules as a simplistic connection table, with the inherent advantages of ease of handling and storage. In the context of virtual screening, chemical databases to be screened are often initially represented by canonicalised SMILES strings that can be filtered and pre-processed in a number of ways, resulting in molecules that occupy similar regions of chemical space to active compounds of a therapeutic target. A wide variety of software exists to convert molecules into SMILES format, namely, Mol2smi (Daylight Inc.), MOE (Chemical Computing Group) and Babel (Openeye Scientific Software). Depending on the algorithm employed, the atoms of a SMILES string defining a molecule can be ordered differently. Upon conversion to 3D coordinates they result in the production of ostensibly the same molecule. In this work we show how different permutations of a SMILES string can affect conformer generation, affecting reliability and repeatability of the results. Furthermore, we propose a novel procedure for the generation of conformers, taking advantage of the permutation of the input strings--both SMILES and other 2D formats, leading to more effective sampling of conformation space in output, and also implementing fingerprint and principal component analyses step to post process and visualise the results.

Algorithms↗

Characteristics of anterior cruciate ligament injuries in Australian football.

Anterior cruciate ligament (ACL) injuries are the most costly injuries in football at both professional and amateur levels (Orchard J, Seward H, McGivern J, Hood S. Intrinsic and extrinsic risk factors for anterior cruciate ligament injury in Australian footballers. Am J Sports Med 2001;29:196-200.). In this study video analysis of 34 ACL injuries in Australian football was performed to investigate the causes of these injuries. Factors that may have contributed to the cause of the injury were analysed, rated and reported. The factors analysed were: type of manoeuvre, direction the knee 'gave way', running speed, knee angle, cutting angle and if the player was accelerating or decelerating. The majority of the injuries analysed occurred in non-contact situations (56%). Of these 37% occurred during sidestepping manoeuvres, 32% in landing, 16% land and step, 10% stopping/slowing and 5% crossover cut manoeuvres. Ninety-two percent of the non-contact injuries occurred at extended knee angles of 30 degrees or less, which is also commonly known to place stress on the ACL and reduce the protective role of hamstrings. Over half (54%) of non-contact injuries occurred whilst decelerating. It would be expected that greater speed and angle cut too would increase the frequency of ACL injury. The results could not confirm this with most injuries occurring at running speeds of slow jogging to running and equal number of injuries occurred at cutting to angles of the ranges 15-45 degrees and 45-75 degrees. These results give greater understanding into potential causes or contributors of ACL injury and information to assist in the development of knee injury prevention programs.

Anterior Cruciate Ligament Injuries↗

A kinematic and kinetic analysis of walking after total knee arthroplasty with and without patellar resurfacing.

BACKGROUND: Clarification of the indications for patellar resurfacing in total knee arthroplasty (TKA) is still necessary. Few studies of adequate power have evaluated functional differences between total knee arthroplasty with and without patellar resurfacing, in particular walking gait. This study aimed to identify clinically relevant differences in knee kinematic or kinetic parameters during level walking between total knee arthroplasty with and without patellar resurfacing, after controlling for pre-surgery gait parameters. METHODS: Kinematic and kinetic gait analysis of level walking was performed on 34 subjects (41 knees) before and 12-18 months after total knee arthroplasty with patellar resurfacing performed randomly. Linear regression analysis was used to examine the influence of patellar resurfacing upon gait variables whilst controlling for the corresponding pre-surgery measure. FINDINGS: The pre-surgery value was a moderate to strong significant predictor of all post-surgery temporal-spatial and kinetic gait parameters (p < 0.001-0.008), and most kinematic parameters (p < 0.001-0.066). The addition of patellar resurfacing to the regression models did not improve the predictive power in any case. Only one parameter, knee flexion at heel-strike, displayed a difference near statistical significance between total knee arthroplasty with and without patellar resurfacing (10 degrees versus 7 degrees respectively, p = 0.023). INTERPRETATION: Pre-surgery gait patterns are an important determinant of post-surgery gait. There are no clinically relevant differences in walking gait between total knee arthroplasty performed with or without patellar resurfacing, using the Profix design.

Aged↗

Oncology exploration: charting cancer medicinal chemistry space.

Approaches for the experimental determination of protein-ligand molecular interactions are reliant on the quality of the compounds being tested. The application of large, randomly designed combinatorial libraries has given way to the creation of more-focused 'drug-like' libraries. Prior to synthesis, we wish to screen the potential compounds to remove undesired chemical moieties and to be within a required range of physiochemical properties. We have used a principal-component analysis (PCA) computational approach to analyze the 3D descriptor space of active and non-active (hit-like) cancer medicinal chemistry compounds. We define hit-like those molecules passing the unmodified OpenEye FILTER program. Our analysis indicates that these compounds occupy quite different regions in space. Cancer-active compounds exist in a much greater volume of space than generic hit-like space and most of them fail the commonly applied filters for orally bioavailable drugs. This is of great significance when designing orally bioavailable cancer target drugs.

Antineoplastic Agents↗

Estrogen receptors: molecular interactions, virtual screening and future prospects.

Identification of the Estrogen Receptor (ER) as a key mediator of the proliferation of breast cancer, and its involvement in pathways leading to osteoporosis and coronary heart disease, has resulted in a surge to discover and design compounds with the ability to modulate its actions (SERMs). Concurrently, a dramatic increase in the number of crystal structures of the ER has led to a more in depth understanding of the governing mechanisms involved in ER modulation. Entwining computational techniques with the availability of 3D structural data, has allowed not only the rational design of potent inhibitors of the ER, but also its incorporation in Virtual Screening (VS) in the search for novel chemotypes that can modulate the ER. An important initial step in the VS process is to filter towards molecules that occupy similar chemical space to a set of known actives prior to docking. We illustrate through Principal Component Analysis (PCA) of 145 descriptors the region of chemical space antiestrogens occupy compared with 'drug-like' space. We also review all available studies involving validation of several docking algorithms utilizing the ER, ultimately focusing on analysis of Enrichment (E) rates and False Positive (FP) rates to illustrate the successes attributed to each docking algorithm. Finally, we relate the recent discovery of non-genomic mechanisms of the ER and subsequently present a model involving a recently identified alternative, second binding-pocket of the ER in our laboratory through cavity analysis that suggests how the same receptor can invoke these, 'classical' and rapid responses concurrently.

Algorithms↗

Antiestrogenically active 2-benzyl-1,1-diarylbut-2-enes: synthesis, structure-activity relationships and molecular modeling study for flexible estrogen receptor antagonists.

The nonsteroidal antiestrogen drug tamoxifen is the endocrine treatment of choice for estrogen receptor positive breast cancer, while the related estrogen receptor antagonist raloxifene is an effective therapeutic intervention for osteoporosis. We report the development of a series of hydroxylated 2-benzyl-1,1-diarylbut-2-enes containing a flexible core scaffold structure differing from the 1,1,2-triarylethylene typical of tamoxifen analogues. In this novel structure, a benzylic methylene group acts as a flexible hinge linking the aryl ring C and the ethylene group. The target products were synthesized using a McMurry coupling (titanium tetrachloride/zinc mediated) procedure. In this study, introduction of hydroxyl, ether and ester substitution on ring C was explored in an attempt to correlate possible metabolic activation in Ring C with antiproliferative activity. These Ring C substituted products showed potent antiproliferative activity against the MCF-7 human breast cancer cell line. The compounds were also shown to have high binding affinity for the estrogen receptor (IC(50) values in the low nanomolar range) together with up to 17 fold selectivity for ERalpha/beta. Some compounds demonstrated antiestrogenic activity in the Ishikawa cells at 40 nM without estrogenic stimulation. The products also displayed a pro-apoptotic effect in MCF-7 cells in a flow cytometry based assay. In a computational study, docked structures of the active compounds were compared with the X-ray crystal structures for the complexes of ERalpha with 4-hydroxytamoxifen and ERbeta with raloxifene. The novel ligands are predicted to bind to the ERalpha and ERbeta in an antiestrogenic orientation, with expected differences obtained in the alignment of the benzylic ring C within the ligand binding domain.

Antineoplastic Agents↗

A hypothesis for the function of braking forces during running turns.

We examined the functional role of braking forces observed when humans execute turning maneuvers. Deceleration caused by braking forces contributes to changing the movement direction of the center of mass (COM) and maintaining constant velocity. We argue that braking forces also prevent over-rotation of the body about the vertical axis during maneuvers. We analyzed data from sidestep and crossover cuts at average initial running velocities of 3 m s(-1). Absent braking, lateral forces would result in body rotations 1.4-3 times the change in COM movement direction, causing the orientation of the body to be substantially mis-aligned with the direction of movement at the end of the step. A simple model based on the hypothesis that body rotation should match COM deflection can explain 70% of the variance in braking forces employed during running turns.

Biomechanical Phenomena↗

Structure-activity studies with high-affinity inhibitors of pyroglutamyl-peptidase II.

Inhibitors of PPII (pyroglutamyl-peptidase II) (EC 3.4.19.6) have potential applications as investigative and therapeutic agents. The rational design of inhibitors is hindered, however, by the lack of an experimental structure for PPII. Previous studies have demonstrated that replacement of histidine in TRH (thyrotropin-releasing hormone) with asparagine produces a competitive PPII inhibitor (Ki 17.5 microM). To gain further insight into which functional groups are significant for inhibitory activity, we investigated the effects on inhibition of structural modifications to Glp-Asn-ProNH2 (pyroglutamyl-asparaginyl-prolineamide). Synthesis and kinetic analysis of a diverse series of carboxamide and C-terminally extended Glp-Asn-ProNH2 analogues were undertaken. Extensive quantitative structure-activity relationships were generated, which indicated that key functionalities in the basic molecular structure of the inhibitors combine in a unique way to cause PPII inhibition. Data from kinetic and molecular modelling studies suggest that hydrogen bonding between the asparagine side chain and PPII may provide a basis for the inhibitory properties of the asparagine-containing peptides. Prolineamide appeared to be important for interaction with the S2' subsite, but some modifications were tolerated. Extension of Glp-Asn-ProNH2 with hydrophobic amino acids at the C-terminus led to a novel set of PPII inhibitors active in vitro at nanomolar concentrations. Such inhibitors were shown to enhance recovery of TRH released from rat brain slices. Glp-Asn-Pro-Tyr-Trp-Trp-7-amido-4-methylcoumarin displayed a Ki of 1 nM, making it the most potent competitive PPII inhibitor described to date. PPII inhibitors with this level of potency should find application in exploring the biological functions of TRH and PPII, and potentially provide a basis for development of novel therapeutics.

Amino Acid Sequence↗

Kinematic and electromyography analysis of submaximal differences running on a firm surface compared with soft, dry sand.

Kinematic and electromyography (EMG) aspects of running on a firm surface and on soft, dry sand were studied to elucidate mechanisms contributing to the higher energy cost (EC) of sand running. Eight well-trained males (mean VO(2max) 64.3+/-8.6 ml.kg(-1).min(-1)) performed barefoot running trials on a firm surface (wooden floor) and on a soft, dry sand surface (track dimensions 8.8 mx60 cm; depth 13 cm) at 8 and 11 km.h(-1). Kinematic and EMG data were collected simultaneously using an integrated six-camera 50 Hz VICON motion analysis system, an AMTI force-plate and a 10-channel EMG system. Running at 8 km.h(-1) on sand resulted in a greater (P<0.05) stance time (t(s)) compared with the firm surface. At 11 km.h(-1), sand running resulted in a greater stance-to-stride ratio (P<0.005), a shorter stride length (SL) (P<0.05), and a greater cadence (P<0.001) compared with the firm surface values. Hip and knee flexion at initial foot contact (IFC), mid-support (MS) and flexion maximum were greater (P<0.001) running on sand compared with firm surface values at 8 and 11 km.h(-1). Over duration of stride, Hamstring (semimembranosus and biceps femoris) EMG was greater running on sand compared with the firm surface at 8 (P<0.001) and 11 (P<0.05) km.h(-1). During the stance phase in the 8-km.h(-1) trials, EMG in the Hamstrings (P<0.001), Vastii (Vastus lateralis and Vastus Medialis) (P<0.02), Rectus femoris (Rec Fem) (P<0.01) and Tensor Fascia Latae (Tfl) (P<0.0001) were greater than the firm surface measures. During stance in the 11-km.h(-1) trials, Tfl EMG was greater (P<0.02) running on sand compared with the firm surface. At IFC and MS, Hamstrings' EMG was greater on sand at both running speeds (P<0.001). For the Vastii (P<0.02), Rec Fem (P<0.0001) and Tfl (P<0.0001) muscles, the EMG at MS running on sand at both speeds was greater than the firm surface values. The increased EC of running on sand can be attributed in part to the increased EMG activation associated with greater hip and knee range of motion compared with firm surface running.

Adult↗

Neuromuscular biomechanical modeling to understand knee ligament loading.

PURPOSE: This article examines our use of EMG-driven neuromuscular biomechanical models to study how muscles stabilize the knee. EMG can be used to establish which activation patterns are used by people for knee stabilization. However, it does not reveal the effectiveness of these patterns. The EMG-driven models provide quantitative comparisons of the effectiveness of the different knee-stabilizing activation patterns. METHODS: Subjects performed static tasks and common sporting maneuvers that challenged knee joint stability. EMG, joint posture and motion, and external forces and moments were measured during these tasks. These data were used to calibrate the EMG-driven neuromuscular biomechanical model. We then used the model to predict the role of muscles in supporting varus and valgus moments at the knee. RESULTS: We found specific muscle activation patterns to support varus and valgus moments. The most potent activation pattern to stabilize the knee is when the hamstrings or quadriceps are required to generate flexion or extension moments, respectively. The next most effective knee-stabilizing pattern is cocontraction of the hamstring and quadriceps. The small biarticular muscles at the knee provided the least support of varus and valgus moments. In the sporting tasks, sidestepping was found to place the anterior cruciate ligament at high risk of injury. We found that the muscles are the main defense against knee ligament injuries in these tasks. CONCLUSION: Traditional biomechanical and neurophysiological methods have shown that there are specific activation patterns used to stabilize the knee. By also using the EMG-driven neuromuscular biomechanical model, we have shown how effective muscles are in stabilizing the knee. This modeling method provides a new tool to understand knee joint stabilization.

Biomechanical Phenomena↗

Estimation of muscle forces and joint moments using a forward-inverse dynamics model.

PURPOSE: This paper presents a forward dynamic neuromusculoskeletal model that can be used to estimate and predict joint moments and muscle forces. It uses EMG signals as inputs to the model, and joint moments predicted are verified through inverse dynamics. The aim of the model is to estimate or predict muscle forces about a joint, which can be used to estimate the corresponding joint compressive forces, and/or ligament forces in healthy and impaired subjects, based on the way they activate their muscles. METHODS: The estimation of joint moments requires three steps. In the first step, muscle activation dynamics govern the transformation from the EMG signal to a measure of muscle activation--a time-varying parameter between 0 and 1. In the second step, muscle contraction dynamics characterize how muscle activations are transformed into muscle forces. The final step requires a model of the musculoskeletal geometry to transform muscle forces to joint moments. Each of these steps involves complex, nonlinear relationships. RESULTS: An application is provided to demonstrate how this model can be used to study the forces in the healthy ankle during dynamometer trials and during gait. The model-predicted estimates of joint moment were found to match experimentally determined values closely. CONCLUSION: Neuromusculoskeletal models that use EMG as inputs can be employed to accurately estimate joint moments. The muscle forces predicted from these models can be used to better understand tissue loading in joints, and to provide in vivo estimates of tensile ligament forces and compressive cartilage loads during dynamic tasks. This tool has great potential for aiding in the study of injury mechanisms in sports.

Ankle↗

Synthesis, structure-activity relationships and antagonistic effects in human MCF-7 breast cancer cells of flexible estrogen receptor modulators.

Estrogen receptors are therapeutic intervention targets for diseases such as osteoporosis and breast cancer with both tamoxifen and raloxifene established as clinical estrogen receptor antagonists. We report a series of novel selective estrogen receptor modulators (SERMs) whose structures are based on a flexible core scaffold differing from the triphenylethylene of tamoxifen analogues through the insertion of a benzylic methylene group as a flexible spacer between the aryl ring C and the ethylene group. A facile synthesis of the target compounds utilises the titanium tetrachloride/zinc mediated McMurry coupling reaction. Successive introduction onto the parent scaffold of hydroxyl functional groups afforded a series of increased potency ligands for the ER - essentially exploring the predicted in vivo metabolic activation of such aromatic SERM ligands. This second generation compound series demonstrated high antiproliferative potency against the MCF-7 human breast cancer cell line, with low cytotoxicity. High ER binding affinity (IC50 20 nM) together with up to 12 fold ERalpha/beta selectivity was also observed. In addition, the compounds displayed antiestrogenic effects at 40 nM when evaluated in the Ishikawa cell line with little estrogenic stimulation. Representative ligands were shown to be pro-apoptotic in human MCF-7 cells in a FACS based assay. Comparison of the docked structure obtained for the most active compound with the X-ray crystal structure reported for the complex of ERalpha and 4-hydroxytamoxifen, predict that these ligands bind in an antiestrogenic manner with some differences being observed in the benzylic Ring C orientation, as expected. This work further demonstrates the tolerance of the estrogen receptor towards flexible modulators.

Antineoplastic Agents↗

Benzoxepin-derived estrogen receptor modulators: a novel molecular scaffold for the estrogen receptor.

We present and examine the efficacy of a novel benzoxepin-based scaffold for modulation of the human estrogen receptor. Receptor tolerance of this new molecular scaffold is examined through presentation of experimentally determined antiproliferative effects on human MCF-7 breast tumor cells and measured binding affinities. The effect of functional group substitution on the benzoxepin scaffold is explored through a brief computational structure-activity relationship investigation with molecular simulation.

Antineoplastic Agents↗

Gait selection in the ostrich: mechanical and metabolic characteristics of walking and running with and without an aerial phase.

It has been argued that minimization of metabolic-energy costs is a primary determinant of gait selection in terrestrial animals. This view is based predominantly on data from humans and horses, which have been shown to choose the most economical gait (walking, running, galloping) for any given speed. It is not certain whether a minimization of metabolic costs is associated with the selection of other prevalent forms of terrestrial gaits, such as grounded running (a widespread gait in birds). Using biomechanical and metabolic measurements of four ostriches moving on a treadmill over a range of speeds from 0.8 to 6.7 m s(-1), we reveal here that the selection of walking or grounded running at intermediate speeds also favours a reduction in the metabolic cost of locomotion. This gait transition is characterized by a shift in locomotor kinetics from an inverted-pendulum gait to a bouncing gait that lacks an aerial phase. By contrast, when the ostrich adopts an aerial-running gait at faster speeds, there are no abrupt transitions in mechanical parameters or in the metabolic cost of locomotion. These data suggest a continuum between grounded and aerial running, indicating that they belong to the same locomotor paradigm.

Animals↗

On the molecular pathology of neurodegeneration in IMPDH1-based retinitis pigmentosa.

Retinitis pigmentosa (RP), the hereditary degenerative disease of the photoreceptor neurons of the retina, probably represents the most prevalent cause of registered blindness amongst those of working age in developed countries. Mutations within the gene encoding inosine monophosphate dehydrogenase 1 (IMPDH1), the widely expressed rate-limiting enzyme of the de novo pathway of guanine nucleotide biosynthesis, have recently been shown to cause the RP10 form of autosomal dominant RP. We examined the expression of IMPDH1, IMPDH2 and HPRT transcripts, encoding enzymes of the de novo and salvage pathways of guanine nucleotide biosynthesis, respectively, in retinal sections of mice, the data indicating that the bulk of GTP within photoreceptors is generated by IMPDH1. Impdh1(-/-) null mice are shown here to display a slowly progressive form of retinal degeneration in which visual transduction, analysed by electroretinographic wave functions, becomes gradually compromised, although at 12 months of age most photoreceptors remain structurally intact. In contrast, the human form of RP caused by mutations within the IMPDH1 gene is a severe autosomal dominant degenerative retinopathy in those families that have been examined to date. Expression of mutant IMPDH1 proteins in bacterial and mammalian cells, together with computational simulations, indicate that protein misfolding and aggregation, rather than reduced IMPDH1 enzyme activity, is the likely cause of the severe phenotype experienced by human subjects. Taken together, these findings suggest that RP10 may represent an attractive target for therapeutic intervention, based upon a strategy combining simultaneous suppression of transcripts from normal and mutant IMPDH1 alleles with supplementation of GTP within retinal tissues.

Animals↗

Scaffold hopping in de novo design. Ligand generation in the absence of receptor information.

We report here the de novo generation of chemotypes and scaffolds for the estrogen receptor, without use of the receptor structure in the assembly phase. Through use of ligand superpositions or a single bound conformation of a known active, a pseudoreceptor can be generated as a design envelope, within which novel structures are readily assembled. Many of these structures have high similarity to known chemotypes. Scaffold hopping is readily achieved within this pseudoreceptor, indicating the advantages of such an approach in discovery research.

Benzene Derivatives↗

The effect of tightly bound water molecules on the structural interpretation of ligand-derived pharmacophore models.

The importance of the consideration of water molecules in the structural interpretation of ligand-derived pharmacophore models is explored. We compare and combine results from recently introduced methods for bound-water molecule identification in protein binding sites and ligand-superposition-based pharmacophore derivation, for the interpretation of ligand-derived pharmacophore models. In the analysis of thymidine kinase (HSV-1) and poly (ADP-ribose) polymerase (PARP), the concurrent application of both methods leads to an agreement in the prediction of tightly bound water molecules as key pharmacophoric points in the binding site of these proteins. This agreement has implications for approaching binding site analysis and consensus drug design, as it highlights how pharmacophore-based models of binding sites can include interaction features not only with protein groups but also with bound water molecules.

Ligands↗